The App. Package, M48, Gene Expression is a CD-ROM containing protocols for purification of total RNA or mRNA from cells and tissues, and for RNA cleanup using MagAttract RNA M48 Kits on the BioRobot M48. These protocols provide operating commands for the instrument and allow easy setup through comprehensive, on-screen instructions. Protocols are available for cell or tissue samples, total RNA, mRNA, or total nucleic acid purification, and RNA cleanup.

The App. Package, M48, Gene Expression CD-ROM contains protocols for purification of total RNA or mRNA from cells and tissues, and for RNA cleanup using MagAttract RNA M48 Kits on the BioRobot M48. Using these protocols and kits on the instrument allows purification of high-quality RNA for consistent analysis of gene expression (see figure "Consistent analysis of relative gene expression").

Principle

MagAttract technology combines the speed and efficiency of silica-based RNA purification with the convenient handling of magnetic particles. The BioRobot M48 performs all steps of sample preparation in a procedure that can be scaled up or down. Up to 48 samples, in multiples of 6, are processed in a single run.

Procedure

MagAttract technology combines the speed and efficiency of silica-based RNA purification with the convenient handling of magnetic particles. Following the automated MagAttract RNA M48 procedure, nucleic acids are purified from lysates in one step through their binding to the silica surface of the particles in the presence of a chaotropic salt. The particles are separated from the lysates using a magnet, and DNA is removed by treatment with RNase-free DNase. The magnetic particles are washed, and RNA is eluted in the buffer provided. Elution volumes may vary from 100 µl to 200 µl. Samples are processed on the BioRobot M48 in 1.5 ml and 2.ml sample tubes. Up to 48 samples in increments of 6 samples can be processed per run.

Protocols on the App. Package, M48, Gene Expression, V1.0

Sample type

Amount

Kit

Nucleic acid

Cultured cells

≤ 1 ml

MagAttract RNA Cell Mini M48 Kit

Total RNA

Enzymatic reaction

100 µl

MagAttract RNA Cell Mini M48 Kit

Total RNA

Tissue

≤ 10 mg

MagAttract RNA Tissue Mini M48 Kit

Total RNA

Cultured cells

≤ 1 ml

MagAttract RNA Cell Mini M48 Kit

Total nucleic acids

Tissue

≤ 10 mg

MagAttract RNA Tissue Mini M48 Kit

Total nucleic acids

Cultured cells

≤ 1 ml

MagAttract Direct mRNA M48 Kit

mRNA

Cultured cells

≤ 2 ml

MagAttract Direct mRNA M48 Kit

mRNA

Applications

The high-quality RNA obtained using MagAttract RNA M48 Kits and the App. Package, M48, Gene Expression is suited for use in many applications, such as: